Movie 4.

Vacuolar or cytosolic death of SPN is dictated by Ply expression. Live fluorescence imaging of GFP-Ply expressing engineered SPN strain following infection in mCherry-Gal8 expressing A549s. In S4 Movies, a SPN is tracked which expressed low amount GFP while in S5 Movie a high GFP expressing variant was monitored. Imaging was performed in Laser scanning confocal microscope starting at 30 min p.i. Total 30 z-stacks were collected for both S4 and S5 movies, out of which MIP of stacks 5-19 (S4 movie) and 7-20 (S5 movie) were shown in the movies. S4 Movie depicts that low GFP expressing SPN containing PCVs were marked with Gal8 and degraded within these structures. S5 Movie shows that high GFP expressing SPN, first associated with Gal8, got detached from these structures and then degraded without further being associated with Gal8. Time interval, 15 min; scale bar, 5 μm.